LOCUS       sequence-374281-        9659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BiFC plasmid for N-terminal tagging of genes with yEmVenus (aa 
            173–238).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9659)
  AUTHORS   Subotic A, Swinnen E, Demuyser L, De Keersmaecker H, Mizuno H, 
            Tournu H, Van Dijck P
  TITLE     A Bimolecular Fluorescence Complementation Tool for Identification 
            of Protein-Protein Interactions in Candida albicans.
  JOURNAL   G3 (Bethesda). 2017 Oct 5;7(10):3509-3520. doi: 
            10.1534/g3.117.300149.
  PUBMED    28860184
REFERENCE   2  (bases 1 to 9659)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1534/g3.117"; 
            journalName: "G3 (Bethesda)"; date: "2017-10-5- 5"; volume: "7"; 
            issue: "10"; pages: "3509-3520"
COMMENT     Sequence Label: BiFC3
FEATURES             Location/Qualifiers
     source          1..9659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4622..4924
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4803..4822
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5273..6067
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5273..6067
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5327..5346)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5937..5956
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      6267..6276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      8890..9478
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9379..9398
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9632..9649
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttaaga acatttctgt accgcaaatg
      361 tatcgacttt ttcgatggtg tgttattgtt gttttgttgc gaaatataat agcttctgct
      421 ttatatcctt tgtgttgatc cttcccctcc tttcatactt gtaaaattac aattggtatt
      481 ttcaaccaat attaccacac gtcatgcact aactcaaatg ggcaaaaaaa agagagatag
      541 gaattacgtc attttattat gtttttacgt ggactagaag gttaagataa tttctggtta
      601 ttttgttgat aattgagttt tgttcgacct ccaccaatct ttaaactctc gttgagtatt
      661 cattttaatg attttttgaa atcagaaggg gtgtttctaa cccaggattg ttttattgag
      721 aggttgaaac atcaatgtgt tagtttattg atttgggttg gtttaataga aaaatatcag
      781 taggtgttca ataattgtag tttattttct tggctaaaat tgttgcgaaa aattgacaaa
      841 ataataacaa aaagtatttt agatgctaaa acataatagt aggaatatct acctgaacca
      901 tttacattca acaactggca agaaccttcc taataatttg caagagtaat cacaattaaa
      961 ccaaaatcaa agtttaccgt gtatatagtt ggacagaaag ctgtcacatt ggtttaatac
     1021 atagttgaat atgtatgcat gaaacaacaa caatacaatt acagctcatg atccttcaat
     1081 taatttcaac attcaaacaa cataacacct tgtgtaaatg aggttatgag tacatgtgaa
     1141 caacaaatct tatgactaaa ctcgggtcgt gtacttttga cgattgagtc atttttacca
     1201 attaggcaat taatttttgc taaattttca ttaaccgcgc tggccatccc aacaaacaat
     1261 ataaaagctg acaacgaaaa aaaaaaaaaa aaaaactcca attctgaaaa atttttcttc
     1321 ttcttcatac taaacaaaac aactataaag ataatcaatt aataatgtca caacaaccta
     1381 gtgaaaataa attatggggt ggtcgattca ctggcgccac tgatccatta atggatttat
     1441 acaatgcttc attaccttat gataaagtta tgtatgatgc agatttaact ggtactaaag
     1501 tatatactca aggattgaac aaattaggat taattaccac tgaagaattg aatctgattc
     1561 atcaaggttt agaacaaatt agacaagaat ggcatgataa taaatttatt attaaagctg
     1621 gtgatgaaga tattcatact gctaatgaac gtagattagg agaaatcatt gggaaaaaca
     1681 tttctggtaa agttcatact ggtagatcca gaaatgatca agttgctact gatatgagaa
     1741 tttttgttcg tgaatcatta cttaatttaa gtaagatttt acatcaattc attacagcaa
     1801 ttcttgaacg agctcataaa gaaattgatg tattaatgcc tggttatact catttacaaa
     1861 aagctcaacc tattagatgg gctcattggt taagttcata tgcaacttat ttcactgaag
     1921 attataaacg tttacaagaa atcattactc gtgttaatca atctccatta ggatctggtg
     1981 ctttagctgg tcatccctac ggtattgatc gtgaattttt agctaaagga ttaggatttg
     2041 atggtgttat tgggaattct ttaactgcag tttctgatcg tgattttgtt gttgaaagtt
     2101 tattttggag tacattattt atgaatcata tttctcgatt ttcagaagat ttaattattt
     2161 attctagtgg tgaatttgga tttataaagt tagctgatgc ttactctact ggttcatcat
     2221 taatgccaca aaagaaaaat cctgattcat tagaattatt acgtgggaaa agtggcagag
     2281 tgtttggtca gttatctggg tttttaatgt caattaaatc cattccatca acctataata
     2341 aagatatgca agaagataaa gaaccattat ttgatgcatt gactaccgta gaacattcaa
     2401 ttttaattgc tactggtgtg atttctactt tactgattga taaacaaaat atggaaaaag
     2461 ctttaactat ggatatgtta gctactgatt tagcagatta tttagttaga aaaggtgttc
     2521 catttagaga aactcatcat atttctggtg aatgtgttag aaaagctgaa gaagaaaaat
     2581 tgtcgggaat tgaccaatta tcatttgaac aatttcaaca aattgattca cgttttgaaa
     2641 aagatgttat ggaaactttt gattttgaag ctagtgtgga aagaagagat gctattggtg
     2701 gtactgctaa gagcgctgtt ttaaaacagt tagaaaattt gaaatcaatt ttaaattaat
     2761 ttataaataa tcatatagta ttcaatgttt tgttttgttt tttttttttt ttttatggaa
     2821 atgtacaaac agtatataat caattaatag aaagataccc tgtattccat tgtatcgcct
     2881 ttgggatgtc tataatacca actttcaaga atgagaaatt aaacaacaat ttctttcacc
     2941 aatcaactcg tctatattca ataaactcat tctggtcttt tcaaatattt aatgttatcc
     3001 tgatcaaact tacaaatcaa aaatgtatca aaatcattca ttagttcttc ttcttttgtt
     3061 atctttgtca ctatatttct ctccatccag accaaaaata cttccattgt gaagatcgtc
     3121 tgaatcataa tcagcaacta cctcttccat ttcagagaat tcagattttt caaaataggt
     3181 atcacgggcc aagtacacac aagacaaacg atacgagtgc tgggatattt ctttcaatgt
     3241 atcactaaat tcgttgagat ttaaagagag ccctttgact gctttcatta ctttcctcac
     3301 atcatgagca tccacattca aatttcctgt agccctttat tcttagtttt cttttcttat
     3361 tcttttcttc gtacattgaa atcacaaatg gtgatatcaa acgaaggtca cactgactta
     3421 tgtctaatac ctccaatgat gatactggta acttaaccag gtctactctt ttcaaacgat
     3481 ttctcattag tgtcaattta cgtaggttct tcaacctttc aaacgatgca aatagctttt
     3541 gtaagggcga attctgcaga tatccatcac actggcggcc ggccgctcga gcatgcatct
     3601 agagggccct tgaagaattg gaagaagtta gtagttaaaa aaacaatggg aaagatttaa
     3661 tctttttttc ttttttcaaa aatttttaac cgctgatgat atattaatct ctctccacca
     3721 ctctctttaa acaccacaat tattaataat aagatatctc tcctactagt tatttgttag
     3781 taagtatatt gttgattgat tggttgggtt aaaaattttt tttttcataa agaaaataat
     3841 gaaagtgaga aaaatttaaa ccccacatta catactcaac acaacactac acacacgcat
     3901 ttaacaatca tcatcagttg aaatatattt tacaaacaat tacgattgtt taatgaaaac
     3961 tatttctgtt gttattatta tttaaccttt tttttcttgt tgggttgacc ctttttctat
     4021 taactatcta actaccacat aacatttgaa gttgaactat taaaccttaa tttgcggccg
     4081 caaatctaat ctactttctt cttcttctac cctatttatt ctgaaatggg gggggggggg
     4141 ggtgggaact ttaggcgtgt taaatttgtc acatctgttc tcacttgctt ctattcgtat
     4201 aattttaatt gtcttgtgag atcaatcttg ttcttgttcg agttttttct tgctttgatt
     4261 aactactatt attattagta tgcaatgctg tttattacca tttcataact tatattataa
     4321 ctcattgatt tcaatacctg ctcaacaatg agtcgttatt attactgtgc tctgtattct
     4381 gtactgctgc acgtacgtta tgtccattga agacaagcaa agaaagacca acattcaatt
     4441 gtatctgctt accagagttg tttacctctg ttaccaatat ttggaaatta attttttggt
     4501 ggtggacgtt tcttcctaaa acacctagat gtgctagtat aattgcacaa ccagcacgca
     4561 cactgcgtct ggccggtcct ggactacatt ttgtctctaa cattcgtagc tgcacgtacg
     4621 gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca
     4681 gagtgatatt attgacacgc cggggcgacg gatggtgatc cccctggcca gtgcacgtct
     4741 gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg
     4801 gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc
     4861 tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat
     4921 ataaatgtca ggcatgagat tatcaaaaag gatcttcacc tagatccttt tcacgtagaa
     4981 agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac
     5041 aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata
     5101 gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc
     5161 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
     5221 atggcgcagg ggatcaagct ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     5281 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5341 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5401 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     5461 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5521 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5581 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5641 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5701 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5761 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     5821 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5881 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5941 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6001 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6061 cttctgaatt attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     6121 cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6181 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6241 tgcttcaata atagcacgtg aggagggcca ccatggccaa gttgaccagt gccgttccgg
     6301 tgctcaccgc gcgcgacgtc gtctattcca agcctgtggc tttttttggt gattcacaat
     6361 acataaattg gcaaatacct cttctataga ctttaacctt tgatgaaatg gtatggataa
     6421 accacttttg tggtcagggt atatatattt ttattccaat ctggccttta taaattcaaa
     6481 tttccaaggg gactctgcta aaccttctga tataactata actctaataa tggctgtaaa
     6541 aagtgtgaaa aaaagatgat attcacaaaa aagataaagg ggatttcata ttgggtatct
     6601 agatttaaac atcgtatttc aacaatgaaa ctagaatcaa tacgaaatcc agggcacaac
     6661 tgtgttcatt cccttgggaa ttatactgag taatgggaag aatgtgcaaa aaatacccgt
     6721 gtcttatata atttccattt gtgtagggct taaaagtgga caacttcagg ttacgaaacc
     6781 acgaaaatgt ctaaacgttt gatcaaattc gtaactcatg gaaaatcaca gcaacaaatt
     6841 aaggatggat atatccggta cttacaaata aaaacactaa agaagtataa attttgaact
     6901 gaaattctat agaaaatctt tttttttggg taaaattatg cgattgtggc tcatagtaac
     6961 gtaatcatac agttcaacaa acatcatttt gtcaaaggat ttctaatcct attctttttc
     7021 tacttggctt ttgagtatat atatatatat atatacatcc tttgttcatc atcatgttcg
     7081 attgaatttt tttcttggtt gttattagat tgtctacttg agtttttctt ggtgcaaatt
     7141 gtggtgtttt tctaacaaat ttttcaacat ttaaagtatt tctggaactt ttctacaagg
     7201 aacgacatag atccattaat gcgcccctct aaaacaatac ccataatgtc ttcttaagta
     7261 tgcatctgct tataaatgtt tctggattta ctttagttag aatgtcgaca cgacaatgaa
     7321 ctattgctgt agatcacgtg cgatcactga attatgagtt tctttttatt tttctttttg
     7381 ggcttttttt tttgcaattt tttttttcca cctttgccaa ttgcgtagtc acatataatg
     7441 tatacaattt ttgcaactct gacactataa ttgatccaat aataaagtat ataatagatc
     7501 atcaagtata cgtaatctcc ccaaaaaaaa aattgcataa aataaaatct ataaaccttt
     7561 tttagatttt tagttgttcc ttttttcttt ctattcttca acttccaagt gttgtcactt
     7621 tcttcctgtt aactatattt aaaagtaaat accctcccca gaaaactccg gaatggatgg
     7681 tggtgttcaa ttagctgacc attatcaaca aaatactcca attggtgatg gtccagtctt
     7741 gttaccagac aaccattact tatcctatca atctaaatta tccaaagatc caaacgaaaa
     7801 gagagaccac atggtcttgt tagaatttgt tactgctgct ggtattaccc atggtatgga
     7861 tgaattgtac aaagctagcg gaggtggagg ttctggtgga ggtggttcaa gcctagggac
     7921 ccgggggcgc gcctaaacgc gtagtactta agatcctgaa gacgggatcg ttcgaagagt
     7981 gaaattctgg aaatctggaa atctggtttt gtattcttgt tattcttctt tttgttatta
     8041 catatataac ttgttacttt tttaaaaaaa tctttgttta ttttataaat atataaaact
     8101 aaatttaaga aaaagagaaa aatgttttat ttgagagatt gaaattttac ttgaatttag
     8161 cttagctttt ataaagtatt attatgtaaa aaaacaaaac aaatatacat taaaaagtta
     8221 agactataaa atagccaccc aaggcatttc tatatcttgt tgttgttgtt ttcatcttct
     8281 gtatcagagg aacttatttt attattttcg tcacgggtat tttctcttgt ttgatgattc
     8341 atcccattca ttccatcata aaattctaga aggaccacct ttgattgtaa atagtaataa
     8401 ttaccaccct tatttaatta tttatttaac ttatttattt atttattata catatataca
     8461 aatctaataa agtgaaaatc tcccccttca caccggtgtg gtccgggacg acgtgaccct
     8521 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     8581 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     8641 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     8701 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
     8761 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     8821 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     8881 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     8941 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     9001 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     9061 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     9121 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     9181 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     9241 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     9301 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     9361 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     9421 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     9481 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     9541 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     9601 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
